| Bone is changing dynamically by environment all the livetime,one of the reason is stress.So,the reconstruction of alveolar bone is becoming one of the most important research field in Prosthodontics.Since osteoblasts are the main cells of bone reconstruction.The cultivation of osteoblasts is becoming the needed means.The research of osteoblast under stress was concentrared in mechanical strain at one time.However, the study of bone in physiological condition recently indicate :the morphological changes of bone is tittle,especially compared whith the study in vitro.So,the fluid shear stress become the main reason of stress resently.How to feel the chang of stress in osteoblast?How to transfer thesignal in osteoblast?How to transfer the signal to osteoclast?There are so many question in the study.The integrins can feel the change of environment.But the effect of fuid shear stress on integrins is remain unkown.In the same time,the effect of fluid shear stress on OPG/OPGL is not clear.This study is centralized in the two field.The purpose of the study is to reveal,how the osteoblast feel the chang of stress ? How the the signal transfer into the osteoblast?And how the signal which transfer to osteoclast change?The major work done under my study is:1. The cultivation of osteoblast from the digestion of calvariaObjective:Cultivation the osteoblast from calvaria of rat.The process can help to establish reliable cell cultivate method and finding the best timing to observe the cell by providing biological effects of osteoblast. Method:Taking the calvaria of rat under asepsis,cutting into pieces.Using pancreatic enzyme and collagen enzyme I to digestion.Using ALP to dye,and observe under microscope,and count the cells .Result and Conclusion:The digestion of calvaria can establish reliable osteoblast.The cells shows very specific characteristic with positive result from ALP dye.The best time of using of osteoblast is 3d-6d.2. Establish the model of osteoblast under fluid shear stress in vitroObjective:Using the device of parallel plate system to makesteady fluid shear stress to osteoblast.To observe the morphological changes of osteoblast and to find the best range of fluid shear stress to osteoblast. Method:Taking osteoblast under fluid shear stress in 6-60 dyn/cm2.Observe the morphological changes under microscope and SEM and TUNNEL dye. Result and Conclusion:The osteoblast changed from a polygonal to a spindle shape under fluid shear stress.. The damage of osteoblast happens above 36 dyn/cm2.The best range of fluid shear stress is 6-36 dyn/cm2.6 dyn/cm2,12 dyn/cm2, 18 dyn/cm2, 36 dyn/cm2 become the standard of fluid shear stress using in study.3. The effect of fluid shear stress on integrin P 1 > a vObjectiverTo study the effect of fluid shear stress on integrin PU a v,search the signal transduction pathway of osteoblast under fluid shear stress. Method:Add fluid shear stress 6 dyn/cm2,12 dyn/cm2,18 dyn/cm2 and 18 dyn/cm2 to osteoblast in 15min ,30min and 60min.The results are detected by RT-PCR. Result and Conclusion: The integrin P K a v increasd under fluid shear stress 6,12,18dyn/cm2.The biggest value occurd in 60 min and 18 dyn/cm2.Under 36 dyn/cm2 fluid shear stress,the integrin PI, ay decreased.The result is consistent with the morphological changes.And the EMC maybe the signal passway of osteoblast under fluid shear stress.4. The effect of fluid shear stress on OPG/OPGLObjective:To investigate the effect of fluid shear stress onOPG/OPGL.And search the theory of bone reconstuction. Method:Add fluid shear stress 6 dyn/cm2,12 dyn/cm ,18 dyn/cm2 and 18 dyn/cm2 to osteoblast in 15min ,30min and 60 min.The results are detected by RT-PCR. Result and Conclusion: The OPG/OPGL increasd under fluid shear stress 6,12,18dyn/cm2. Under 36 dyn/cm2 fluid shear stress,OPG/OPGL decreased.Under 18 dyn/cm2,OPG and OPGL are correspond.OPGL is bigger than OPG under 36 dyn/cm2. This means the construction of bone is workly under aptitude stress,and bone damage become under big stress. |
PDF Full Text Request